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National Chung Hsing University

12 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Electrotaxis Studies of Lung Cancer Cells using a Multichannel Dual-electric-field Microfluidic Chip
Hsien-San Hou 1, Hui-Fang Chang 1, Ji-Yen Cheng 1,2,3,4,5
1Research Center for Applied Sciences, Academia Sinica, 2Institute of Biophotonics, National Yang-Ming University, 3Biophotonics & Molecular Imaging Research Center (BMIRC), National Yang-Ming University, 4Department of Mechanical and Mechatronic Engineering, National Taiwan Ocean University, 5Ph.D. Program in Microbial Genomics, National Chung Hsing University

Many microfluidic devices have been developed for use in the study of electrotaxis. Yet, none of these chips allows the efficient study of the simultaneous chemical and electric-field (EF) effects on cells. We developed a polymethylmethacrylate-based device that offers better-controlled coexisting EF and chemical stimulation for use in electrotaxis research.

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Bioengineering

A Microfluidic Platform for High-throughput Single-cell Isolation and Culture
Ching-Hui Lin 1,2, Hao-Chen Chang 1,2, Chia-Hsien Hsu 1,2,3
1Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Taiwan, 2Tissue Engineering and Regenerative Medicine, National Chung Hsing University, 3Institute of NanoEngineering and MicroSystems, National Tsing Hua University

Here, we present a protocol for isolating and culturing single cells with a microfluidic platform, which utilizes a new microwell design concept to allow for high-efficiency single cell isolation and long-term clonal culture.

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Engineering

Probing C84-embedded Si Substrate Using Scanning Probe Microscopy and Molecular Dynamics
Mon-Shu Ho 1, Chih-Pong Huang 2, Jyun-Hwei Tsai 3, Che-Fu Chou 1, Wen-Jay Lee 3
1Department of Physics and Institute of Nanoscience, National Chung Hsing University, 2Metallurgy Section, Materials & Electro-Optics Research Division, National Chung-Shan Institute of Science and Technology, 3National Center for High-Performance Computing

This paper reports the nanomaterial fabrication of a fullerene Si substrate inspected and verified by nanomeasurements and molecular dynamic simulation.

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Bioengineering

Designing Microfluidic Devices for Studying Cellular Responses Under Single or Coexisting Chemical/Electrical/Shear Stress Stimuli
Tzu-Yuan Chou 1, Yung-Shin Sun 2, Hsien-San Hou 3, Shang-Ying Wu 1, Yun Zhu 1, Ji-Yen Cheng 3, Kai-Yin Lo 1
1Department of Agricultural Chemistry, National Taiwan University, 2Department of Physics, Fu-Jen Catholic University, 3Research Center for Applied Sciences, Academia Sinica

Micro-fabricated devices integrated with fluidic components provide an in vitro platform for cell studies mimicking the in vivo micro-environment. We developed polymethylmethacrylate-based microfluidic chips for studying cellular responses under single or coexisting chemical/electrical/shear stress stimuli.

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Environment

Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level
Chong-Yu Ko 1, Yue-Wen Chen 1, Yu-Shin Nai 1
1Department of Biotechnology and Animal Science, National Ilan University

Herein we present a method to feed pesticide contaminated food to both an individual honey bee and a beehive colony. The procedure evaluates the pesticide effect on individual honey bees by in vivo feeding of basic larval diet and also on the natural condition of beehive colony.

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Genetics

Transient Expression of Foreign Genes in Insect Cells (sf9) for Protein Functional Assay
Ju-Chun Chang 1, Se Jin Lee 2, Jae Su Kim 2, Chung-Hsiung Wang 3, Yu-Shin Nai 1
1Department of Biotechnology and Animal Science, National Ilan University, 2Department of Agricultural Biology, College of Agriculture Life Sciences, Chonbuk National University, 3Department of Entomology, National Taiwan University

This protocol describes a heat shock-induced protein expression system (pDHsp/V5-His/sf9 cell system), which can be used for either expressing foreign proteins or evaluating the anti-apoptotic activity of potential foreign proteins and their truncated amino acids in insect cells.

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Environment

Use of Autometallography to Localize and Semi-Quantify Silver in Cetacean Tissues
Wen-Ta Li 1, Bang-Yeh Liou 1, Wei-Cheng Yang 2, Meng-Hsien Chen 3, Hui-Wen Chang 1,2, Hue-Ying Chiou 4, Victor Fei Pang 1,2, Chian-Ren Jeng 1,2
1Graduate Institute of Molecular and Comparative Pathobiology, National Taiwan University, 2School of Veterinary Medicine, National Taiwan University, 3Department of Oceanography and Asia-Pacific Ocean Research Center, National Sun Yat-sen University, 4Graduate Institute of Veterinary Pathobiology, National Chung Hsing University

A protocol is presented to localize Ag in cetacean liver and kidney tissues by autometallography. Furthermore, a new assay, named the cetacean histological Ag assay (CHAA) is developed to estimate the Ag concentrations in those tissues.

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Bioengineering

Establishing Single-Cell Based Co-Cultures in a Deterministic Manner with a Microfluidic Chip
Cheng-Kun He 1,2, Ya-Wen Chen 3, Ssu-Han Wang 3, Chia-Hsien Hsu 1,2
1Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, 2Ph.D. Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, 3National Institute of Cancer Research, National Health Research Institutes

This report describes a microfluidic chip-based method to set up a single cell culture experiment in which high-efficiency pairing and microscopic analysis of multiple single cells can be achieved.

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Bioengineering

Electric-Field-Induced Neural Precursor Cell Differentiation in Microfluidic Devices
Hui-Fang Chang 1, Shih-En Chou 1, Ji-Yen Cheng 1,2,3,4
1Research Center for Applied Sciences, Academia Sinica, 2Institute of Biophotonics, National Yang Ming Chao Tung University, 3Department of Mechanical and Mechatronic Engineering, National Taiwan Ocean University, 4College of Engineering, Chang Gung University

In this study, we present a protocol for the differentiation of neural stem and progenitor cells (NPCs) solely induced by direct current (DC) pulse stimulation in a microfluidic system.

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Biology

Isolation and Selection of Entomopathogenic Fungi from Soil Samples and Evaluation of Fungal Virulence against Insect Pests
Yao-Chia Liu 1, Nian-Tong Ni 1, Ju-Chun Chang 1, Yi-Hsuan Li 1, Mi Rong Lee 2, Jae Su Kim 2,3, Yu-Shin Nai 1
1Department of Entomology, National Chung Hsing University, 2Department of Agricultural biology, Jeonbuk National University, 3Department of Agricultural Convergence Technology, Jeonbuk National University

Here we present a protocol based on the mealworm (Tenebrio molitor)-bait system that was used for isolating and selecting entomopathogenic fungi (EPF) from soil samples. An effective conidia number (ECN) formula is used to select high stress tolerant EPF based on physiological characteristics for pest microbial control in the field.

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Bioengineering

Inactivation of Pathogens via Visible-Light Photolysis of Riboflavin-5′-Phosphate
Chien-Wei Cheng 1, Shwu-Yuan Lee 2, Tang-Yu Chen 1, Jeu-Ming P. Yuann 1, Chi-Ming Chiu 1, Shiuh-Tsuen Huang 3,4, Ji-Yuan Liang 1
1Department of Biotechnology, Ming Chuan University, 2Department of Tourism and Leisure, Hsing Wu University, 3Department of Science Education and Application, National Taichung University of Education, 4Department of Soil and Environmental Sciences, National Chung Hsing University

Here, we present a protocol to inactivate pathogenic bacteria with reactive oxygen species produced during photolysis of flavin mononucleotide (FMN) under blue and violet light irradiation of low intensity. FMN photolysis is demonstrated to be a simple and safe method for sanitary processes.

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Biology

Assessment of Aphidicidal Effect of Entomopathogenic Fungi against Parthenogenetic Insect, Mustard Aphid, Lipaphis erysimi (Kalt.)
Cheng-Ju Yang 1, Yu-Shin Nai 1
1Department of Entomology, National Chung Hsing University

This protocol presents an optimized detached-leaf bioassay system for evaluating the effectiveness of entomopathogenic fungi (EPF) against the mustard aphid (Lipaphis erysimi (Kalt.)), a parthenogenetic insect. The method outlines the data collection process during Petri dish experiments, enabling researchers to consistently measure the virulence of EPF against mustard aphids and other parthenogenetic insects.

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